The overall aim of this project is to study the renal excretion and metabolism of methylamines related to choline and the effect of certain agents on choline transport and metabolism in the kidney. Choline is a precurser of betaine, acetylcholine and phosphatidylcholine and may be degraded in the gut to the various methylamines. The effect of chronic ethanol administration on choline transport and metabolism will be examined using the isolated perfused rat kidney. During ethanol addiction and withdrawal plasma choline levels will be measured. The effect of deanol (dimethylaminoethanol) on the renal excretion and metabolism of choline (trimethylaminoethanol) will be studied using isolated perfused rat kidney and the Sperber technique in hens. These same biological systems will be used to study the renal transport and metabolism of tri-, di- and mono-methylamines during administration of the substrates at various concentrations. Blood levels of choline and the methylamines will be measured in rats on high and low choline diets and compared with blood levels obtained in rats in which uremia will be induced by uretural ligation. Choline will be measured by a choline kinase assay using 32P-ATP and forming 32P-phosphorylcholine. The methylamines will be measured by gas chromatography. Subcellular localization of the renal N-oxidase will be determined by incubation of cellular fractions obtained through centrifugation techniques using the conversion of 14C-trimethylamine to 14C-trimethylamine N-oxide as a model. The ability of the renal N-oxidase system to participate in the metabolism and excretion of nicotinamide, deanol, meperidine and amphetamine will be studied during perfusion of the isolated rat kidney and infusion into the renal portal circulation in hens. In those experiments where metabolites are measured the 14C-substrate will be administered. Electrophoresis and chromatography will be used to isolate the 14C-components in the biological fluids followed by quantitation using scintillation counting of radioactivity.